黄芩苷通过ROS/MAPK/HIF-1α信号通路颉颃双酚A诱导的鸡巨噬细胞凋亡和程序性坏死  

Baicalin antagonizes bisphenol A-induced apoptosis and necroptosis in chicken macrophages via the ROS/MAPK/HIF-1αsignaling pathway

作　　者：曹舒婷 王天佑 曲丹杨 徐世文[1] CAO Shuting;WANG Tianyou;QU Danyang;XU Shiwen(College of Veterinary Medicine,Northeast Agricultural University,Harbin 150030,China)

机构地区：[1]东北农业大学动物医学学院,哈尔滨150030

出　　处：《黑龙江畜牧兽医》2025年第2期86-94,共9页Heilongjiang Animal Science And veterinary Medicine

基　　金：黑龙江省自然科学基金联合引导项目(LH2023C024)。

摘　　要：为了探究黄芩苷(BAI)通过活性氧(ROS)/丝裂原活化蛋白激酶(MAPK)/缺氧诱导因子-1α(HIF-1α)信号通路对双酚A(BPA)诱导的鸡巨噬细胞(HD11)凋亡和程序性坏死的颉颃作用,试验采用不同质量浓度的BPA和/或BAI处理HD11细胞24 h,通过CCK-8法测定BPA对HD11细胞的半数抑制浓度,并筛选出BAI颉颃BPA的最适剂量。将HD11细胞分为对照组(正常培养)、BAI组(加入确定的BAI最适剂量)、BPA组(加入确定的BPA最适剂量)和BAI+BPA组(加入确定的BPA和BAI最适剂量),常规培养24 h后收集细胞,检测细胞凋亡和程序性坏死情况及氧化应激指标,并检测细胞凋亡、程序性坏死和ROS/MAPK/HIF-1α信号通路相关基因和蛋白的表达。结果表明:BPA对HD11细胞的半数抑制浓度为(45.55±0.09)μg/mL,BAI颉颃BPA的最适剂量为0.89μg/mL。与对照组相比,BPA组细胞内ROS相对荧光强度显著增加(P<0.05),而与BPA组相比,BAI组和BPA+BAI组细胞内ROS相对荧光强度显著降低(P<0.05)。与对照组相比,BPA组总抗氧化能力及谷胱甘肽过氧化物酶、过氧化氢酶、超氧化物歧化酶活性显著降低(P<0.05),而丙二醛含量显著升高(P<0.05);与BPA组相比,BAI组和BPA+BAI组总抗氧化能力及谷胱甘肽过氧化物酶、过氧化氢酶、超氧化物歧化酶活性显著升高(P<0.05),而丙二醛含量显著降低(P<0.05)。与对照组相比,BPA组ROS/MAPK/HIF-1α信号通路相关因子(JNK、HIF-1α、P38和ERK)表达量显著升高(P<0.05),除了凋亡因子Bcl-2的表达量显著降低(P<0.05)外,其余细胞凋亡相关因子(P53、BAK和Cytc)和程序性坏死相关因子(RIPK1、RIPK3和MLKL)表达量显著升高(P<0.05);而与BPA组相比,BAI组和BPA+BAI组ROS/MAPK/HIF1α信号通路相关因子(JNK、HIF-1α、P38和ERK)表达量显著降低(P<0.05),除了凋亡因子Bcl-2的表达量显著升高(P<0.05)外,其余细胞凋亡相关因子(P53、BAK和Cytc)和程序性坏死相关因子(RIPK1、RIPK3和MLKL)表达量显著降低(P<0.05)。说�In order to explore the antagonizing effect of baicalin(BAI)on bisphenol A(BPA)-induced apoptosis and necroptosis in chicken macrophages(HD11)via reactive oxygen species(ROS)/mitogen activated protein kinase(MAPK)/hypoxia inducible factor-1α(HIF-1α)signaling pathway,in this test,HD11 cells were treated with different concentrations of BPA and/or BAI for 24 h,and the half inhibitory concentrations of BPA on HD11 cells were determined by CCK-8 method,and the optimal dose of BAI antagonizing BPA was screened.HD11 cells were divided into control group(normal culture),BAI group(with the addition of the determined BAI optimal dose),BPA group(with the addition of the determined BPA optimal dose),and BAI+BPA group(with the addition of the determined BPA and BAI optimal doses);cells were collected in a routine culture for 24 h.Apoptosis and necroptosis,oxidative stress indexes and the expression of genes and proteins related to apoptosis,necroptosis and ROS/MAPK/HIF-1αsignaling pathway were detected.The results showed that the half inhibitory concentration of BPA on HD11 cells was(45.55±0.09)μg/mL,and the optimal dose of BAI antagonizing BPA was 0.89μg/mL.Compared with the control group,the relative fluorescence intensity of intracellular ROS in the BPA group was significantly increased(P<0.05),while the relative fluorescence intensity of intracellular ROS in the BAI group and BPA+BAI group was significantly decreased compared with the BPA group(P<0.05).Compared with the control group,the total antioxidant capacity and the activities of glutathione peroxidase,catalase and superoxide dismutase in the BPA group were significantly decreased(P<0.05),while the content of malondialdehyde was significantly·increased(P<0.05).Compared with the BPA group,the total antioxidant capacity,glutathione peroxidase,catalase and superoxide dismutase activities in the BAI group and BPA+BAI group were significantly increased(P<0.05),while the malondialdehyde content was significantly decreased(P<0.05).Compared with the control group,th

关 键 词：双酚A(BPA) 黄芩苷(BAI) 鸡巨噬细胞(HD11) 细胞凋亡 程序性坏死 ROS/MAPK/HIF-1α 

分 类 号：S859[农业科学—临床兽医学]